Analysis of substrate metabolism for an immobilized CYP2C9 construct: A potential bioreactor application

BIOT 263

Lance Wollenberg, lwollenb@mix.wvu.edu1, Jarod L. Kabulski1, Timothy S. Tracy2, David Lederman3, and Peter M. Gannett1. (1) Department of Pharmaceutical and Pharmacological Sciences, West Virginia University, 1 Medical Center Drive, Morgantown, WV 26505, (2) Department of Experimental and Clinical Pharmacology, University of Minnesota, Minneapolis, MN 55455, (3) Department of Physics, West Virginia University, Morgantown, WV 26506
Essential in the metabolism of xenobiotics are the heme-containing enzymes, the cytochrome P450s (CYP). We have demonstrated a method to immobilize CYP2C9 on gold. The metabolic activity of this construct was verified using Δ-9-tetrahydrocannibinol (THC), a high turnover CYP2C9 substrate. Here we report on the metabolic competency of immobilized CYP2C9 using 1) model CYP2C9 substrates, including non-steroidal anti-inflammatory drugs, which often show lower substrate turnover rates in comparison to THC, and 2) the effect of various effector molecules known to accelerate model substrate metabolism in vitro in our immobilized CYP2C9 constructs. The results for dapsone (effector) on flurbiprofen (substrate) metabolism parallel solution data. Additional examples will be discussed. Our results show that the immobilized enzyme activity parallels the in vitro activity. Therefore this system could be used in bioreactors for metabolite production, creating a practical tool for drug development. (Supported by WVEPSCoR STEM program and NIH GM063215 and GM069753.)
 

Poster Session
5:30 PM-7:30 PM, Wednesday, August 22, 2007 BCEC -- Exhibit Hall - B2, Poster

Division of Biochemical Technology

The 234th ACS National Meeting, Boston, MA, August 19-23, 2007